Product Class: Restriction Endonuclease

DpnI
recombinant time saver CutSmart_long dil_B 37 80 Heat CpG Epi

 

dpnIcutsite

Product Introduction

  • DpnI cleaves only when its recognition site is methylated.
  • Cleavage of mammalian genomic DNA is blocked by overlapping CpG methylation.

Having supplied restriction enzymes to the research community for over 40 years, NEB has earned the reputation of being the leader in enzyme technologies. Working continuously to be worthy of that distinction, NEB strives to develop enzymes of the highest purity and unparalleled quality.

Bioz Badge Exists : True
Catalog # Size Concentration
R0176S 1,000 units 20,000 units/ml
R0176L 5,000 units 20,000 units/ml
Need a custom/large volume order? Contact Us

Product Information

Description


Product Source

An E. coli strain that carries the DpnI gene from Diplococcus pneumoniae G41 (S. Lacks).

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
Product Categories:
Restriction Enzymes for Epigenetic Analysis,
Epigenetic Analysis,
Restriction Endonucleases C G
,
time-saver-qualified-restriction-enzymes Products
Applications:
Site Directed Mutagenesis,
Site Directed Mutagenesis,
Restriction Enzymes for Epigenetics
,
Fast Cloning: Accelerate your cloning workflows with reagents from NEB
,
Restriction Enzyme Digestion

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to digest 1 µg of pBR322 DNA (dam methylated) in 1 hour at 37°C in a total reaction volume of 50 µl.

Reaction Conditions

1X CutSmart® Buffer
Incubate at 37°C

1X CutSmart® Buffer
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
100 µg/ml BSA
(pH 7.9 @ 25°C)

Activity in NEBuffers

NEBuffer™ 1.1: 100%
NEBuffer™ 2.1: 100%
NEBuffer™ 3.1: 75%
CutSmart® Buffer: 100%

Diluent Compatibility

Storage Conditions

10 mM Tris-HCl
300 mM NaCl
1 mM DTT
0.1 mM EDTA
500 µg/ml BSA
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

80°C for 20 min

Methylation Sensitivity

dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Blocked by Overlapping

Product Notes

  1. DpnI cleaves only when its recognition site is methylated. DNA purified from a dam+ strain will be a substrate for DpnI.
  2. Blocked by overlapping CpG methylation.

Protocols, Manuals & Usage

Protocols

  1. Optimizing Restriction Endonuclease Reactions
  2. Optimizing Restriction Endonuclease Reactions
  3. Restriction Digest Protocol
  4. Double Digest Protocol with Standard Restriction Enzymes

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. Will DpnI cleave hemimethylated DNA?
  2. What's the difference between DpnI, DpnII, MboI, and Sau3AI?
  3. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
  4. Why is my Restriction Enzyme not cutting DNA?
  5. Why do I see a DNA smear on an agarose gel after a restriction digest?
  6. Why do I see additional DNA bands on my gel after a restriction digest?
  7. How many nucleotides do I have to add adjacent to the RE recognition site in order to get efficient cutting?
  8. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
  9. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?

Troubleshooting

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.